Milk Extracellular Vesicles and Their Functional Potential in Osteoblast Activity: A Cross‐Species Comparison for Nutraceutical Development

ABSTRACT Milk is a complete food that plays a fundamental role in supporting bone health with high content of calcium and protein. Recent research highlights the significance of milk‐derived extracellular vesicles (mEVs) with emerging roles in modulating physiological processes, including bone remodeling. This study reports a comparative analysis of mEVs isolated from raw milk of three ruminant species: bovine, goat, and sheep. Extracellular vesicles (EVs) are characterized for size distribution, particle concentration, and protein composition. Functional relevance of each type of mEVs is evaluated on cell proliferation and osteogenic differentiation of murine fibroblasts model. Results reveal significant interspecies differences in both the structural and biological characteristics of mEVs. EVs derived from each milk type exhibit distinct effects on MC3T3‐E1 viability, supporting the idea that mEVs have a beneficial role in cell growth and proliferation and induce osteogenic differentiation. Additionally, the microbial DNA content carried within mEVs from each ruminant species harbors distinct and structured microbial DNA profiles. Goat mEVs are enriched in beneficial commensals, whereas sheep samples reflect consistent yet mixed communities, and bovine mEVs exhibit environmental signatures and greater variability. Overall, these findings highlight the species‐specific potential of mEVs, their compositional diversity, and potential functions of the microbial cargo. Furthermore, this study highlights mEVs as functional food components and opens new avenues for developing targeted dietary strategies leveraging mEVs to support personalised medicine in bone health and prevent or manage conditions such as osteoporosis.